Dispensing device and system for biological products

ABSTRACT

A dispensing system has a sterilizable chamber. At least one robotic arm assembly and a dispensing device are within the sterilizable chamber. The dispensing device comprising a first dispensing element and a second dispensing element. The first dispensing element is connectable to the second dispensing element The at least one robotic assembly is configured to move the dispensing device. An external control device is connected to the at least one robotic arm assembly and is configured to control the at least one robotic arm assembly. The first dispensing element is configured to dispense a structural material and the second dispensing element is configured to dispense a biological material.

BACKGROUND 1. Field of the Invention

The application relates to a handheld dispensing device, as well as asystem provided therewith, and a dispensing system comprising asterilizable chamber.

2. Description of the Related Art

Three-dimensional (3D) bioprinting is the process of employing 3Dprinting technologies using materials such as cells and supportingcomponents in order to create e.g. cell patterns and living tissues,wherein the cell function and viability are preserved within the printedconstruct. Recently, 3D bioprinting has begun to incorporate theprinting of scaffolds, i.e. structures providing support for the cells.

Current state-of-the-art solutions for 3D bioprinting are large 3Dprinting machines with components that move over a printing tray fillede.g. with nutrient rich media, potentially causing particle sheddinginto the tray. An example of a single-use biological 3D printer is givenin US 2015/0035206 A1.

Alternatively, small-scale solutions such as pen printers are available.For example, U.S. Pat. No. 8,834,793 B2 discloses a pen capable ofdispensing biological material, whereas U.S. Pat. No. 9,102,098 B2discloses a pen capable of dispensing structural material used forprinting scaffolds. Generally, structural material (e.g. thermoplastic)pen printers and biological material pen printers (such as celldispensers) exist separately. The ARC Centre of Excellence forElectromaterials Science developed a pen that extrudes cell materialinside a biopolymer such as alginate, which is in turn encased in anouter layer of gel material. Both the outer and inner layers arecombined in the pen head as it is extruded. However, the pen must besterilized after every use and it may be cumbersome to comply with therequired sterilization conditions.

There is therefore a need for a system allowing a biological material tobe added to structural material and scaffolding to efficiently andeasily form a plurality of objects under sterile conditions. Inparticular, a handheld device allowing a user to quickly, easily, andcheaply try out multiple designs at the small-scale while checking forstructural material and cell compatibility and optimizing their processprior to e.g. scaling up to a large-scale biological 3D printer isneeded.

One application may be for laboratories and biopharma companies toquickly screen materials, scaffolding designs, and shapes to test theircells and bioactive materials for material compatibility and to optimizetheir process prior to scale-up. The primary application for thebiopharma industry may be for screening of 3D printed cell products forefficacy and toxicity testing prior to use with the single-usebiological 3D printers. This small-scale lab technology may additionallybe applied to the screening the customer's bioactive materials for InVitro Diagnostic (IVD) tests on diagnostic membranes such as the UniSartline of products, printing on biosensors, as well as a variety of custommedical devices.

SUMMARY

According to one aspect, a handheld dispensing device is provided. Thehandheld dispensing device comprises the following: a first dispensingelement configured to dispense a structural material; and a seconddispensing element configured to dispense a biological material; whereinthe second dispensing element is single-use and sterilizable; andwherein the first dispensing element is releasably connectable to thesecond dispensing element.

According to another aspect, a system is provided. The system comprisesthe following: a handheld dispensing device according to the firstaspect; at least one single-use bioreactor; and at least one asepticconnection assembly configured to connect the at least one single-usebioreactor to the handheld dispensing device.

According to a further aspect, a dispensing system is provided. Thedispensing system comprises the following: a sterilizable chamber; atleast one robotic arm assembly and a dispensing device within thesterilizable chamber, wherein the at least one robotic assembly isconfigured to move the dispensing device comprising a first dispensingelement and a second dispensing element, the first dispensing elementbeing releasably connectable to the second dispensing element; and anexternal control device connected to the at least one robotic armassembly and configured to control the at least one robotic armassembly; wherein the first dispensing element is configured to dispensea structural material and the second dispensing element is configured todispense a biological material.

Details of exemplary embodiments are set forth below with reference tothe exemplary drawings. Other features will be apparent from thedescription, the drawings, and from the claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A and 1B shows an example of a handheld dispensing devicecomprising an attachment mechanism.

FIGS. 2A-2D shows an example of a handheld dispensing device comprisinga body provided with at least one sleeve.

FIGS. 3A-3D shows other examples of a handheld dispensing devicecomprising an attachment mechanism.

FIGS. 4A-4C shows examples of a handheld dispensing device comprisingone or more temperature-regulating elements.

FIGS. 5A-5D shows examples of dispensing elements comprising differentmechanisms.

FIG. 6 shows an example of a system comprising a handheld dispensingdevice and a biological material source.

FIG. 7 shows an example of a system comprising a handheld dispensingdevice and two biological material sources.

FIG. 8 shows an example of a system comprising a handheld dispensingdevice, a biological material source, a centrifugation assembly and acrossflow assembly.

FIG. 9 shows an example of an external monitoring setup for themonitoring of a handheld dispensing device.

FIG. 10 shows an example of a dispensing system comprising asterilizable chamber, a dispensing device and a manually-operatedexternal control device.

FIG. 11 shows an example of a dispensing system comprising asterilizable chamber, a dispensing device and a automatically-operatedexternal control device.

DETAILED DESCRIPTION

In the following text, a detailed description of examples will be givenwith reference to the drawings. It should be understood that variousmodifications to the examples may be made. In particular, one or moreelements of one example may be combined and used in other examples toform new examples. In the following, reference numbers with a primedenote the same components as the corresponding plain numbers but in adifferent configuration. For example, 10 denotes a dispensing device ina disconnected state and 10′ denotes a dispensing device in a connectedstate.

FIGS. 1A to 11 show different examples of a dispensing device that cansupply both a structural material and a biological material for additivemanufacturing. In other words, the materials being extruded by thedispensing device can be used to print, i.e. create, 3D objects. A 3Dobject is any object that exists in the three dimensions of space,regardless of the measures of its length, width and depth and the ratiosthereof.

Structural materials encompass materials whose primary purpose is tocarry the loads and/or maintain the juxtaposition of the component partsof a system. Structural materials are load or stress bearing, whereinthe stress can be induced either mechanically, thermally or acombination of both. Structural materials may be used to buildscaffolds, i.e. structures providing support for cells and othermaterials derived from a biological system. The architecture of thescaffold is important for e.g. structural, nutrient transport, andcell-matrix interaction conditions. Examples of structural materials areheated thermoplastics, hydrogels, cellulose, collagen, collagen fibers,bio-paper (and other dissolvable materials), sugars, epoxies, syntheticpolymers.

Biological materials may include materials comprising a biologicalsystem, such as cells, cell components, cell products, and othermolecules, as well as materials derived from a biological system, suchas proteins, antibodies and growth factors. The dispensing device may inaddition dispense other materials related to the biological material,e.g. supportive fluids such as nutrient rich media and metered drugproducts.

The dispensing device described here provides a platform for dispensingstructural materials to form a scaffold or a three dimensional printedobject as well as biological materials that may be dispensed onto thestructural materials to form a printed product. A wide variety ofstructures such as cell structures, biological products, implants,scaffolding structures, diagnostic kits, etc. may be printed.

The dispensing device can utilize a variety of different techniques ofadditive manufacturing whereby materials are built up layer-by-layer toform the 3D products. The dispensing means may include but are notlimited to an extrusion printing head that deposits a solid, fluid, orsemi-solid material in a continuous stream or in droplets, a heatedextrusion printing head that can heat a solid material to help it flowas a fluid where it can solidify once printed onto a platform, and aspray deposition printing head which sprays micronized droplets of amaterial over a specific area within a pattern.

The dispensing device may be a handheld device held directly by anoperator, as shown in FIGS. 1A to 9, or it may be controlled by arobotic control tool, as shown in FIGS. 10 and 11. According to oneexample, the robotic control tool may be manually controlled by anoperator via a communication with a stylus simulant maneuvered by theoperator. According to another example, the robotic control tool may runa pre-programmed operation for printing.

The dispensing device may comprise at least two dispensing elements, thefirst dispensing element configured to dispense the structural materialand the second dispensing element configured to dispense the biologicalmaterial. In some examples, the dispensing device may compriseadditional dispensing elements, e.g. for dispensing material related tothe biological material, for example material required to preserve thebiological material and/or its viability, such as supportive fluids. Theadditional dispensing elements may include, but are not limited to, apipette dispensing element, a fluid dispensing element, a spray coatingdispensing element, a metered drug dispenser, a nutrient rich mediadispenser, a gel dispenser, a sputter coating dispenser, and an adhesivedispensing tool. One or more of the dispensing elements may comprisehydraulic, pneumatic, magnetic, electric, and/or manual drive mechanismsconfigured to control the movement of the material until it is dispensedfrom a dispensing tip. In one example, the drive mechanism may compriseat least one feed motor.

Additionally or alternatively, the dispensing device may comprise one ormore modification tools for modifying the printed object. Themodification tools may include but are not limited to a puncturing tool,a blade, a cutting tool, a rotating screw, a grinding tool, animprinting tool, a stamping tool, and a laser cutter tool.

Exemplarily, the dispensing device may comprise a material selectorswitch to select which dispensing element among the plurality ofdispensing elements in the dispensing device should dispense itsmaterial. In other words, the material selector switch may select thematerial to be dispensed by the dispensing device.

The first dispensing element (also referred to as “structural materialdispensing element”) and the second dispensing element (also referred toas “biological material dispensing element”) may be fixedly orreleasably connectable to each other. Similarly, the additionaldispensing elements and/or modification tools may be connectable. Thedispensing elements may be directly connected, e.g. physicallycontacting each other, or indirectly connected, e.g. via a thirdelement. The connection between the dispensing elements may allow power,data (e.g. sensor data, input data) and/or fluid communication betweenthe dispensing elements.

In some examples, the dispensing elements are permanently connected toeach other, in that the dispensing device is formed as a whole. In otherexamples the dispensing elements may be releasably connectable. In otherwords, the dispensing elements may exist in a physically separated stateor in a connected state, and it is possible to switch back and forthfrom one state to the other. The releasable connection may beaccomplished via an attachment mechanism, such as a snap mechanismincluding protrusions and corresponding fitted holes to accommodate theprotrusions. Other attachment mechanisms may comprise but are notlimited to displaceable locking tabs and fitted receiving holders,fasteners and magnets.

In some examples, the dispensing device may comprise barriers betweenthe dispensing elements to prevent undesired effects due to theproximity of the dispensing elements when connected. Exemplarily, thefirst dispensing element may comprise a heating element to heat thestructural material and a barrier (e.g. a thermal barrier) may shieldthe biological material, preventing the heating from negativelyaffecting the biological material in the second dispensing element.

At least the second dispensing element is a sterilizable, single-useelement to prevent cross contamination from one batch of biologicalmaterial to another. A single-use element is a disposable element, i.e.an element that is discarded after being used. The single-use element isconfigured for a one-time use and, after it has been used once, it hasfulfilled its function and may be disposed of. A single-use element isformed from sterilizable materials and helps reduce the risk ofcontaminations because of its disposability.

The sterilizable, single-use biological material dispensing element maybe delivered pre-sterilized, such as by gamma-irradiation, or besterilized by the operator using a validated sterilization method, suchas by autoclaving, prior to use. The sterilizable, single-use biologicalmaterial dispensing element may connect with the current infrastructureof bioreactors, both multi-use and single-use, micro-bioreactors,filters, and other bioprocessing devices through standardized asepticconnectors. For example, the biological material dispensing element mayutilize an aseptic connection to a biological material feed source suchas a bioreactor vessel. The biological material from the feed source mayundergo processing such as filtration, concentration, etc., prior todispensing from the dispensing element.

According to one example, the sterilizable, single-use biologicalmaterial dispensing element may be connected to a multi-use structuralmaterial dispensing element. According to another example, the entiredispensing device comprising the first and second dispensing elementsmay be sterilizable and single-use.

The dispensing device is intended to allow an operator to quickly screenfor structural material compatibility with biological materials. Theoperator is able to easily create a plurality of shapes, designs,structures, scaffolding, and printing procedures to optimize for thebest combination. Essentially this is determining if the material typesand shapes printed are compatible for maintaining or promoting cellgrowth and optimizing the conditions for such growth prior to scaling upactivities to a large-scale biological 3D printer, such as a single-useprinter assembly.

The dispensing device may contain one or more orientation andpositioning sensors or may be positionally tracked by an externalmonitoring device to record the movements, materials dispensed, andconditions. This stored positional file of the recorded movements of thedispensing device may be utilized to replicate a qualified printeddesign from the small-scale dispensing device within a large-scalebiological 3D printer. In other words, the stored information regardingthe method and conditions of dispensing may be used to print with a 3Dprinter the structural materials and biological materials in the exactsame way an operator constructed a three dimensional object using thedispensing device.

FIGS. 1A and 1B shows an example of a handheld dispensing device 10comprising an attachment mechanism. A sterilizable, single-usebiological material dispensing element 50 is directly connected to amulti-use structural material dispensing element 12 using the attachmentmechanism. FIG. 1A is a side view of the dispensing device 10 in adisconnected state, wherein the dispensing device 10 comprises the firstdispensing element 12 and the second dispensing element 50 disconnectedfrom each other.

The structural material dispensing element 12 of the dispensing device10 may comprise a printer body 14 that encloses a plurality ofcomponents. The multi-use structural material dispensing element 12 ofthe dispensing device 10 may utilize a structural material 16, which forexample is a thermoplastic material, such as polylactic acid (PLA),acrylonitrile butadiene styrene (ABS), carbon fiber, low melting pointmetals and/or other low melting point structural materials. In otherexamples, the structural material may consist of but is not limited tohydroxyapatite (HA), collagen, fibrin, hydrogels, chitosan, hyaluronicacid, sugars, gels, powders.

In this example, a length of thermoplastic structural material 16 may beinserted into the top of the multi-use structural material dispensingelement 12 and the dispensing speed is determined by a feed motor 18that has at least one gear 20 to control the movement of thethermoplastic structural material 16. A plurality of feed motors 18 andgears 20 may be utilized to ensure a steady flow of the structuralmaterial 16 through the top of the dispensing element 12 until it isdispensed from a dispensing tip 38 to prevent jamming internally withinthe device.

The structural material 16 may move through an internal tube 22 to aheating element 24 such as an electrically-powered heating element,which is powered through an electrical connection 46. In this example,the heating of the thermoplastic structural material 16 and thedispensing through the narrow dispensing tip 38 may constitute a form ofultra-pasteurization that can kill off micro-organisms within thestructural material 16 even if it is from a non-sterile feedstock. Otherdispensing methods without heating or compression may require asepticconnection to a sterile feedstock prior to use.

An operator may control the temperature of the heating element 24 andthe dispensing of the structural material 16 through a computer controlboard 26 that may contain a processing device 28, a memory storagedevice 30, and a wireless communications device 32. The multi-usestructural material dispensing element 12 may comprise one or aplurality of controlling devices to allow the operator to control theflow of material, e.g. utilizing dispense and retraction buttons 34.Furthermore, a material selector switch 36 may be provided to selectwhich material from a plurality of materials should be dispensed by thedispensing device 10, such as switching from the dispensing of astructural material 16 to the dispensing of a biological material, oncethe second dispensing element 50 is connected to the first dispensingelement 12. Additionally or alternatively a remote input device (notshown) may be utilized, such as a wired or wireless foot pedal, awearable device, or other remote input, to control the flow of materialand the selection of materials to dispense.

The dispensing device 10 may contain in the first dispensing element 12a plurality of positional sensors 44 including but not limited to motionsensors, orientation sensors, gyroscopic sensors, environmental sensors,cameras, microscopic cameras, thermal cameras, depth sensors, ultrasounddevices, magnetometers, accelerometers, proximity sensors, globalpositioning system (GPS) devices, internal measurement units (IMUs) andinternal or external positioning sensors. The information from theorientation and positioning of the dispensing device 10 used by anoperator can be saved into a file, which may be utilized to print out areplica of the structural materials and biological materials in theexact same way as was manually printed utilizing a large-scalesingle-use biological 3D printer setup (not shown). Additionally oralternatively the sensors utilized for positioning may also be utilizedto scan the workspace, the printing tray, and/or the three dimensionalobject prior to, during, and/or after printing. These scans may beutilized to determine the structure or microstructures of the printedmaterials.

The multi-use structural material dispensing element 12 of thedispensing device 10 may use an attachment mechanism 42, e.g. a snapmechanism, to attach to a corresponding attachment mechanism 64 on thesingle-use biological material dispensing element 50. In this example,the dispensing elements 12 and 50 are directly connected. The attachmentmechanism 42 may bring the single-use biological material dispensingelement 50 in communicative connection with the multi-use structuralmaterial dispensing element 12, wherein the connection may establish anelectrical, mechanical, and/or fluid communication. In this example theconnection between the multi-use structural material dispensing element12 and the single-use biological material dispensing element 50 maycontain a thermal barrier 40 to prevent excess heat from the heatingelement 24 to negatively impact the dispensing of the biologicalmaterials.

The single-use biological material dispensing element 50 may beconstructed of sterilizable materials and may be sterilized utilizing avalidated method, such as gamma irradiation, autoclaving, and/orchemical sterilization. For example, the single-use biological materialdispensing element 50 may consist of a printer body 52 that encloses aplurality of components. Furthermore, it may include at least onephysical aseptic connector that may comprise two or more components,such as an OPTA® connector with a tubing to connect to an externalsterile feed source of cells, media, and/or other biological materials,as well as a tubing component 58 that transports the sterile feedmaterial from the aseptic connection to an internal tubing 54.Alternatively the aseptic connection may be made with at least onethermoweldable tubing which can be connected using a Biowelder®thermoweldable tubing sealer (not shown).

The internal tubing 54 runs the length of the printer body 52 until adispensing tip 56 situated at the end opposite the tubing component 58.The dispensing tip 56 may comprise a plurality of shapes to alter thedispensing profile of the dispensed material, such as slow and focusedor sprayed over a defined area or pattern. The dispensing tip 56 mayadditionally contain a restriction device (not shown) that may partiallyclose to restrict the material flow through the tip 56 or completelyclose to stop the flow of all material through the tip 56. Alternativelythe operator may select between a plurality of dispensing tips, withdifferent geometries for different dispensing patterns, which may beselected by moving a carousel wheel containing multiple tipconfigurations (not shown) on the printer body 52 or by attachingpre-sterilized tips (not shown) to the dispensing tip 56.

A vent filter 62, such as an integrity-testable, sterilizing-grade ventfilter, may be utilized to properly vent the internal tubing 54 as itfills with material. The movement of the material through the single-usebiological material dispensing element 50 may be provided by an externalsource such as a pneumatic pressure source, a pump, or an electricmotor. Alternatively, the pump, electric motor, and/or pneumaticpressure source may be internal to the dispensing device 10.

FIG. 1B is a side view of the dispensing device 10′ in a connectedstate. In this example the multi-use structural material dispensingelement 12′ is connected using attachment mechanism 42′, which isattached to the corresponding attachment mechanism 64′ on the single-usebiological material dispensing element 50′. The material selector switch36′ may be moved from a position for selecting the multi-use structuralmaterial dispensing element 12′ to a position for selecting thesingle-use biological material dispensing element 50′, so the biologicalmaterial and/or material related to the biological material may bedispensed by the operator utilizing the material dispensing andretraction buttons. The dispensing device 10′ in the connected state maycontain an ergonomically-molded handgrip on either the materialdispensing element 12′, the single-use biological material dispensingelement 50′, or across both elements for the comfort of the operatorusing the device.

A power cable 48 may be plugged into the electrical connection 46 topower the electric heating element 24, the at least one feed motor 18,or other electronic components. Alternatively, the dispensing device 10may be powered by at least one of a battery that is exemplarilyrechargeable, an alternate wired connection, a powered communicationport, a solar cell, a mechanical power source, an electromechanicalpower source such as a hand crank, or a wireless power source.

The single-use biological material dispensing element 50′ may beconnected to a biological feed source 66 via the at least one asepticconnection 60′. The aseptic connection 60′ may be made using an asepticconnector such as an OPTA® connector and/or other physical asepticconnector, and/or may be achieved by thermowelding a tubing length ofthermoweldable tubing to the feed assembly for a sterile connection withthe biological feed source 66.

FIGS. 2A-2D shows an example of a handheld dispensing device 100comprising a body provided with at least one sleeve 120.

FIG. 2A is a side view of the handheld dispensing device 100 in adisconnected state. The dispensing device 100 comprises a reusablemulti-use structural material dispensing element 102 and the sleeve 120for inserting a sterilizable, single-use biological material dispensingelement 122. The multi-use structural material dispensing element 102 ofthe dispensing device 100 may comprise a printer body 104 that enclosesa plurality of components. In one example, the sleeve 120 may be a partof the printer body 104, so that the dispensing elements 120 and 122 aredirectly connected.

The multi-use structural material dispensing element 102 may utilize astructural material 106 such as a thermoplastic material. In otherexamples the structural material may comprise any of hydroxyapatite(HA), collagen, fibrin, hydrogels, chitosan, hyaluronic acid, sugars,gels, powders.

In this example a length of thermoplastic structural material 106 may beinserted into the top of the multi-use structural material dispensingelement 102 and the dispensing speed may be determined by a feed motorwhich has one or more gears to control the movement of the thermoplasticstructural material 106. The structural material 106 may move through aninternal tube towards a heating element 108, which may heat thethermoplastic structural material 106 and dispense it through a narrowdispensing tip 112.

An operator may control the temperature of the heating element 108 andthe dispensing of the structural material 106 through a computer controlboard 114 that may contain a processing device, a memory storage device,and a wireless communications device. The multi-use structural materialdispensing element 102 may comprise one or a plurality of controllingdevices to allow the operator to control the flow of material, e.g.utilizing dispense and retraction buttons 110 and a material selectorswitch 116, which selects a material from a plurality of dispensingmaterials. Exemplarily, once the single-use biological materialdispensing element 122 is connected to the structural materialdispensing element 102, the material selector switch may switch from thedispensing of the structural material 106 to the dispensing of abiological material.

The dispensing device 100 may contain a plurality of positional sensors118 including but not limited to motion sensors, orientation sensors,gyroscopic sensors, environmental sensors, cameras, depth sensors,magnetometers, accelerometers, proximity sensors, GPS devices, andinternal or external positioning sensors.

The printer body 104 of the multi-use structural material dispensingelement 102 may contain a barrel or sleeve 120 into which the single-usebiological material dispensing element 122 may be inserted. Theconnection between the dispensing elements 102 and 122 via insertioninto the barrel 120 may be further aided by an attachment mechanism,such as a snap mechanism. In this example, the dispensing elements 102and 122 are directly connected. The insertion of the single-usebiological material dispensing element 122 and the optional attachmentto the attachment mechanism may bring the dispensing element 122 incommunicative contact with the multi-use structural material dispensingelement 102. The connection may establish an electrical, mechanical,and/or fluid communication. In this example the connection between themulti-use structural material dispensing element 102 and the single-usebiological material dispensing element 122 may contain a thermal barrierto prevent excess heat from the heating element 108 to negatively impactthe dispensing of the biological materials.

The single-use biological material dispensing element 122 may beconstructed of sterilizable materials and may be sterilized utilizing avalidated method, such as gamma irradiation, autoclaving, and/orchemical sterilization. Exemplarily, the single-use biological materialdispensing element 122 may comprise a printer body that encloses aplurality of components. Furthermore, the dispensing element 122 mayinclude at least one physical aseptic connector 124 that may comprisetwo or more components, such as an OPTA® connector with a tubingcomponent 126 to connect to an external sterile feed source of cells,media, and/or other biological materials, wherein the tubing component126 may transport the sterile feed material from the aseptic connectionto an internal tubing 128. Alternatively the aseptic connection may bemade with at least one thermoweldable tubing that can be connected usinga Biowelder® thermoweldable tubing sealer (not shown).

The internal tubing 128 may run the length of the printer body until adispensing tip 132 at one end opposite the end connected to the tubingcomponent 126. The dispensing tip 132 may comprise a plurality of shapesto alter the dispensing profile of the dispensed material, such as slowand focused or sprayed over a defined area or pattern. A vent filter134, such as an integrity-testable, sterilizing-grade vent filter, maybe utilized to properly vent the internal tubing 128 as it fills withmaterial.

The movement of the material through the single-use biological materialdispensing element 122 may be provided by an external source such as apneumatic pressure source, a pump, or an electric motor. Alternatively,the pump, electric motor, and/or pneumatic pressure source may beinternal to the dispensing device 100.

FIG. 2B is a side view of the dispensing device 100′ in a connectedstate. In this example the single-use biological material dispensingelement 122′ may be inserted into the sleeve 120 and further connectedusing an attachment mechanism. The material selector switch 116′ may bemoved from a position for selecting the multi-use structural materialdispensing element 102′ to a position for selecting the single-usebiological material dispensing element 122′, so the biological materialand/or material related to the biological material may be dispensed bythe operator utilizing the material dispensing and retraction buttons110. The dispensing device 100′ in the connected state may contain anergonomically-molded handgrip on the material dispensing element body102′ for the comfort of the operator using the device.

The single-use biological material dispensing element 122′ may beconnected to a biological feed source 130 via at least one asepticconnection 124′. The aseptic connection 124′ may be achieved bythermowelding a tubing length of thermoweldable tubing to the feedassembly for a sterile connection with the biological feed source. Theaseptic connection 124′ may be made using an aseptic connector such asan OPTA® connector and/or other physical aseptic connector, and/or maybe achieved by thermowelding a tubing length of thermoweldable tubing tothe feed assembly for a sterile connection with the biological feedsource.

The dispensing device 100′ may be powered by a power cable (not shown),a battery, which is exemplarily rechargeable, a powered communicationport, a solar cell, a mechanical power source, an electromechanicalpower source such as a hand crank, or a wireless power source.

FIG. 2C is a side view of a dispensing device 150 in a disconnectedstate. The structural material dispensing element 162 and the biologicalmaterial dispensing element 164 may be similar to the correspondingelements described with reference to views ‘A’ and ‘B’. However, thebody of the structural material dispensing element 162 does not containa sleeve. Rather, in this example, there is a central connection body152 that contains a plurality of barrels or sleeves 154, 156 where theplurality of dispensing elements 162, 164 may be inserted to form acompleted assembly. The dispensing elements 162 and 164 may thus beindirectly connected via the central connection body 152.

A further attachment mechanism may be used to fix the dispensingelements in the sleeves 154, 156. The attachment mechanism may bring theplurality of dispensing elements 162, 164 in communicative connectionwith the central connection body 152, wherein the connection may includeelectrical, mechanical, and/or fluid communication.

In this example the central connection body 152 may be sterilizable andsingle-use. The plurality of dispensing elements 162, 164 connecting tothe central connection body 152 may additionally be sterilizable andsingle-use. The central connection body 152 may contain a thermalbarrier 158 between the plurality of barrels 154, 156 to prevent thermalenergy transfer from negatively affecting the biological materialdispensing element 164. The central connection body 152 may additionallycontain a plurality of controlling elements 160 such as buttons,switches, knobs, and other operator control elements similar to thosedescribed with reference to views ‘A’ and ‘B’.

FIG. 2D is a side view of the dispensing device 150′ where both themulti-use structural material dispensing element 162′ and the single-usebiological material dispensing element 164′, separated by the thermalbarrier 158, are in a connected state and attached to the centralconnection body 152 using an attachment mechanism. The centralconnection body 152 may contain an ergonomically-molded handgrip for thecomfort of the operator using the device.

In this example the multi-use structural material dispensing element162′ may contain a computer control board comprising a processingdevice, a memory storage device, and a wireless communications device.The plurality of controlling elements 160 on the central connection body152 may line up and communicatively attach to the computer control boardelements to receive the inputs of the controlling elements 160.Alternatively the computer control board and other devices may be inpart or in whole integrated into the central connection body 152.

The single-use biological material dispensing element 164′ may beconnected to a biological feed source, such as material originating fromor processed from a multi-use or a single-use bioreactor, via at leastone aseptic connection 166′. For example, the aseptic connection is madeby an aseptic connector such as an OPTA® connector and/or other physicalaseptic connector where two or more components 166′, 168 are connectedtogether. In other examples, the multi-use structural materialdispensing element 102 may require an aseptic connection for somestructural materials, such as hydrogels, hydroxyapatite (HA), collagen,fibrin, chitosan, hyaluronic acid, etc., which may be unable to beheated to extreme temperatures. Since the heating ensures the killing ofall microorganisms in the structural material prior to dispensing, ifheating is not possible, the feed material may require sterilizationprior to connection to the dispensing device 150 to prevent anypotential contamination of the final printed object.

As shown in FIGS. 1A-2D, the structural material dispensing element maygenerally comprise at least one feed motor to control the movement ofthe structural material, at least one internal tubing to contain thestructural material and at least one dispensing tip from which thestructural material is dispensed. Furthermore, the structural materialdispensing element may comprise at least one controlling device (e.g.dispense and retraction buttons and/or material selector switch) tocontrol the flow of the material and select which material must bedispensed when other dispensing elements are connected to the structuralmaterial dispensing element. Additionally or alternatively, thestructural material dispensing element may comprise processing means,such as a computer control board, to control the dispensing process.Exemplarily, the structural material dispensing element may comprise aheating element to heat and melt the structural material, and theprocessing means may be configured to control the temperature of theheating element.

The biological material dispensing element may generally comprise a bodyformed out of sterilizable materials, at least one internal tubing tocontain the biological material, at least one vent filter to properlyvent the internal tubing as it fills with material, and at least onedispensing tip from which the biological material is dispensed.Additionally, the biological material dispensing element may comprise anaseptic connector to a material feed source for the sterile transfer anddispensing of the biological material.

The combination of the biological material dispensing element and thestructural material dispensing element as described in FIGS. 1A-2D showonly one of many possible configurations of a handheld dispensingdevice. FIGS. 3A-3D show other examples of a handheld dispensing devicecomprising an attachment mechanism, wherein a plurality of dispensingelements may be connected and/or an external barrier between connecteddispensing elements may be provided.

FIG. 3A is a side view of a dispensing device 230 where a plurality ofconnectable dispensing elements are in the disconnected state. In thisexample the dispensing elements making up the dispensing device 230 maycomprise a multi-use structural material dispensing element 232, asingle-use biological material dispensing element 262, and a pipetteliquid dispensing element 244, which may be multi-use or single-use. Inother examples, different dispensing elements may be part of thedispensing device 230, wherein the dispensing elements may include butare not limited to spray dispensers, meter drug dispensers, liquidmedium dispensers, gel dispensers, or sputter coating dispensers.Additionally other modification tools to modify the dispensed structuralmaterial and/or the biological material such as a puncturing tool,blade, rotating screw, laser cutter, pressurized sterile air, membranedispensers (for dispensing membranes, fleeces, thin films, andshape-memory polymers), electrospinning fiber/nanofiber dispensers,dispensers for fluorescence or DNA identification tagging, or othertools may be attached to the dispensing device 230 for modifications ofthe printed object. The use of these post-printing modification toolsfor surface modification and the creation of internal pathways withinthe printed three-dimensional structures are described in U.S. patentapplication Ser. No. 14/680,180, which is incorporated herein byreference.

In this example the multi-use structural material dispensing element 232dispenses a structural material 234, which may exemplarily be athermoplastic material. The structural material 234 may move through aninternal tube (not shown) to a heating element (not shown) and bedispensed through a narrow dispensing tip 240. An operator may controlthe temperature of the dispensing of the structural material 234.Further, the operator may control the flow of the material 234 through aplurality of controlling devices such as material dispense andretraction buttons 238 and a material selector switch 236, which selectswhich material from a plurality of materials should be dispensed by thedispensing device 230 when dispensing elements 262 and 244 areconnected.

The multi-use structural material dispensing element 232 may use anattachment mechanism 242, e.g. a snap mechanism, to attach to acorresponding attachment mechanism 258 on the pipette liquid dispensingelement 244.

The pipette liquid dispensing element 244 may comprise a plunger button246, a volume adjustment knob 250, a volume indicator display 252, a tipejector button 248, a tip holder 254, and a disposable pipette tip 256.The pipette liquid dispensing element 244 may contain an internal pistonassembly (not shown) with a barrel, shaft, spring, and O-ring. Thepipette liquid dispensing element 244 may be utilized for dispensing ofa meter volume of a drug or chemical product or for the dispensing ofliquid media or other growth-promoting fluids onto the printed object.

The pipette liquid dispensing element 244 may use an attachmentmechanism 260, e.g. a snap mechanism, to attach to a correspondingattachment mechanism 270 on the single-use biological materialdispensing element 262.

The single-use biological material dispensing element 262 may beconstructed of sterilizable materials and may be sterilized utilizing avalidated method, such as gamma irradiation, autoclaving, and/orchemical sterilization. In this example the single-use biologicalmaterial dispensing element 262 may comprise a printer body whichencloses

A plurality of components. Furthermore, it may include at least onephysical aseptic connector that may comprise two or more components,such as an OPTA® connector with a tubing to connect to an externalsterile feed source of cells, media, and/or other biological materials,as well as a tubing component that transports the sterile feed materialfrom the aseptic connection to an internal tubing (not shown).

The internal tubing (not shown) may run the length of the printer bodyuntil a dispensing tip 268 at one end. The dispensing tip 268 maycomprise a plurality of shapes to alter the dispensing profile of thedispensed material, such as slow and focused or sprayed over a definedarea or pattern. A vent filter 266, such as an integrity-testable,sterilizing-grade vent filter, may be utilized to properly vent theinternal tubing (not shown) as it fills with material. The movement ofthe material through the single-use biological material dispensingelement 262 may be provided by an external source such as a pneumaticpressure source, a pump, or an electric motor. Alternatively a pump,electric motor, and/or pneumatic pressure source may be internal to thedispensing device 230.

FIG. 3B is a side view of the dispensing device 230′ where the pluralityof connectable dispensing elements are in the connected state. In thisexample the multi-use structural material dispensing element 232′, thepipette liquid dispensing element 244′, and the single-use biologicalmaterial dispensing element 262′, are connected utilizing theirrespective attachment mechanisms. The attachment mechanisms may bringthe pipette liquid dispensing element 244′, the single-use biologicalmaterial dispensing element 262′ and the multi-use structural materialdispensing element 232′ in communicative contact with one another,wherein the connection may establish an electrical, mechanical, and/orfluid communication. Between each of the plurality of dispensingelements within the dispensing device 230′ there may be thermal barriers(not shown) to prevent thermal energy transfer from negatively affectingthe dispensing elements 244 and 262. The dispensing device 230′ in theconnected state may contain an ergonomically-molded handgrip on thematerial dispensing element 232′, the pipette liquid dispensing element244′, the single-use biological material dispensing element 262′, oracross all elements for the comfort of the operator using the device.

In the connected configuration of the dispensing device 230′, thematerial selector switch 236′ may change the dispensing elementselected, such as to either the pipette liquid dispensing element 244′or the single-use biological material dispensing element 262′, in orderto allow the operator to control the flow of material utilizing thematerial dispense and retraction buttons 238. The pipette liquiddispensing element 244′ may utilize either the material dispense andretraction buttons 238 or the pipette plunger button 246 on thedispensing element itself, depending on which is more comfortable forthe operator to operate. The dispensing tips 240, 254, 268 of theirrespective dispensing elements are sufficiently spaced to preventoverlapping or spraying onto another element, such as a spray coatingliquid from dispensing element 244′ coming into contact with the heateddispensing tip of the structural material dispensing element 232′.

The single-use biological material dispensing element 262′ may beconnected to a biological feed source 272 via at least one asepticconnection 264′. The aseptic connection 264′ may be made using anaseptic connector such as an OPTA® connector and/or other physicalaseptic connector, and/or may be achieved by thermowelding a tubinglength of thermoweldable tubing to the feed assembly for a sterileconnection with the biological feed source.

FIG. 3C is a side view of a dispensing device 280 where the multi-usestructural material dispensing element 282 is attached to a thermalbarrier 286 to prevent a heating element as discussed in the previousexamples from negatively affecting the biological material within thesingle-use biological material dispensing element 284. The thermalbarrier 286 may be formed in such a way that it provides communicativecontact between the multi-use structural material dispensing element 282and the single-use biological material dispensing element 284 throughthe attachment mechanism (not shown). The thermal barrier 286 may extenddown to the dispensing tips 288, 289 to prevent any negative heatingeffects from the heating element during dispensing of structuralmaterial from dispensing tip 288 and the dispensing of biologicalmaterial from dispensing tip 289.

FIG. 3D is a side view of a dispensing device 290 where the multi-usestructural material dispensing element 292 is attached to a spacerelement 296 to prevent a heating element as discussed in the previousexamples from negatively affecting the biological material within thesingle-use biological material dispensing element 294. The spacerelement 296 may be formed in such a way that it provides communicativecontact between the multi-use structural material dispensing element 292and the single-use biological material dispensing element 294 throughthe attachment mechanism (not shown). The spacer element 296 may onlypartially extend along the lengths of the dispensing elements 292, 294to leave a gap between the dispensing tips 298, 299 where airflow fromthe workspace may prevent any negative heating effects from the heatingelement during dispensing of structural material from dispensing tip 298and the dispensing of biological material from dispensing tip 299.

As discussed with reference to FIGS. 1A to 3D, the structural materialdispensing element may comprise a temperature-regulating element such asa heating element. Additionally or alternatively, othertemperature-regulating elements, such as a cooling element and a warmingelement, may be comprised in the dispensing device in order to regulatethe temperature of the material to be dispensed. In one example,thermoregulation may be accomplished by one or more tubing lines in thedispensing device filled with thermally-regulated fluid. Thethermally-regulated tubing lines may be filled, properly vented, andrecirculated using an external pump and/or pneumatic pressurized source.The temperature regulation device for the fluid may be an externaldevice that heats and/or cools the thermally-regulated fluid within acontainer and recirculates the fluid through the thermally-regulatedtubing lines. In another example, thermoregulation may be accomplishedby means of a single-use chemical temperature-regulating element, whichmay use an exothermic and/or endothermic chemical reaction for asingle-use thermoregulation of the material to be dispensed. FIGS. 4A-4Cshows examples of a handheld dispensing device comprising one or moretemperature-regulating elements.

FIG. 4A is a side view of a single-use dispensing device 300 consistingof a printer body 302 that encloses a plurality of componentsmanufactured from sterilizable materials and may be sterilized utilizinga validated method, such as gamma irradiation, autoclaving, and/orchemical sterilization. In this example, the dispensing device 300 maycomprise a structural material dispensing element that dispensesmaterial out of dispensing tip 312 and a biological material dispensingelement that dispenses material out of dispensing tip 318. In otherwords, the dispensing elements are permanently connected and may beformed together. The single-use dispensing device 300 may contain anergonomically-molded handgrip for the comfort of the operator using thedevice.

The structural material dispensing element of the dispensing device 300may utilize a structural material 304 such as a low melting pointthermoplastic material, which has a lower melting point then the plasticmaterials used for the printer body 302 and other internal componentswithin the dispensing device 300. In other examples the structuralmaterial may consist of hydroxyapatite (HA), collagen, fibrin,hydrogels, chitosan, hyaluronic acid, sugars, gels, powders, or otherstructural materials that do not require heating beyond the meltingpoint temperatures of the plastic materials used for the printer body302 and other internal components within the dispensing device 300.

The structural material 304 may be inserted into the top of thestructural material dispensing element portion of the dispensing device300 and the dispensing speed may be determined by a feed motor (notshown) that has a plurality of gears (not shown) to control the movementof the thermoplastic structural material. In this example in which thewhole dispensing device is single-use, the feed motor (not shown) may bedriven by an external pneumatic/hydraulic pressure source attached to afluid drive and thermoregulation connector 306.

In one example a pneumatic pressure source may be connected to the fluiddrive and thermoregulation connector 306. In this case, the waste air,after running through the feed motor (not shown), may simply bleed thepressure off through the filter 305 in a direction away from the printedobject or the filter may be connected to a length of tubing that removesthe compressed air in excess away from the work area. In anotherexample, a hydraulic pressure source may be connected to the fluid driveand thermoregulation connector 306. In this case, the waste fluidpressure after running through the feed motor (not shown) may use atubing line to drain the compressed fluid away from the work area or torecover the fluid in a container (not shown) where it can be pressurizedand recirculated through the device.

The structural material 304 may move through an internal tube (notshown) to a single-use heating element (not shown), which is describedin more detail below with reference to views FIGS. 4B and 4C.

The single-use biological material dispensing element of the dispensingdevice 300 may be connected to a biological feed source via at least oneaseptic connection 314. A vent filter 316, such as anintegrity-testable, sterilizing-grade vent filter, may be utilized toproperly vent the internal tubing (not shown) as it fills with material.The aseptic connection 314 may be made using an aseptic connector suchas an OPTA® connector and/or other physical aseptic connector, and/ormay be achieved by thermowelding a tubing length of thermoweldabletubing to the feed assembly for a sterile connection with the biologicalfeed source.

An operator may control the dispensing of the structural and biologicalmaterials through dispense and retraction buttons 310 and a materialselector switch 308, that may switch from dispensing the structuralmaterial to dispensing the biological material. In this example thecontrols may regulate the direction and force of the pneumatic/hydraulicpressure that controls the flow of materials within the single-usedevice 300.

FIG. 4B is a side view of a single-use dispensing device 320 comprisinga fluid temperature control mechanism. The single-use dispensing device320 may be a possible embodiment of the single-use dispensing device300.

The single-use dispensing device 320 may comprise a printer body 322that encloses a plurality of components manufactured from sterilizablematerials and may be sterilized utilizing a validated method, such asgamma irradiation, autoclaving, and/or chemical sterilization.

The device 320 may comprise internal recirculation channels within athermal regulating assembly 333 to transfer fluid for heating andcooling elements internal to the dispensing device 320. The individualfluid-conducting elements may be connected through a single connection,such as the fluid drive and thermoregulation connector 306 as shown inview ‘A’, to align and connect each of the individual tubing lines. Thetubing lines may be segregated and insulated from one another to preventthe excess heat or cooling to negatively impact the temperature of anearby temperature controlled tubing line. The tubing lines may compriseat least one of a fluid line 330, a heated tubing line 340, a cooledtubing line 346 and a warm tubing line 366, which will be described inmore detail in the following.

Exemplarily, a structural material dispensing element 324 may comprise astructural material 326, a feed motor 328, a venting filter 332,internal tubing 334, a heating element 336, a cooling element 342, and adispensing tip 348. The feed motor 328 may be driven by an externalpneumatic pressure source attached to the fluid line 330. The waste air,after running through the feed motor 328, may simply bleed the pressureoff through the venting filter 332 in a direction away from the printedobject or the venting filter 332 may be connected to a length of tubingthat removes the excess compressed air away from the work area.

The feed motor 328 may turn a plurality of gears that feed thestructural material 326 into the internal tubing 334 to the heatingelement 336. The heating element 336 may comprise an assembly thatcirculates a heated fluid, such as heated sterile filtered water,glycol, and/or steam, from an external temperature-regulated fluidsource through the heated tubing line 340. The heated tubing line 340may be connected to an internal heating recirculation loop 338 that maybe insulated to protect the internal components and othertemperature-controlled fluid lines from being negatively affected by theelevated temperatures. The heating element 336, the heated tubing line340, and the internal heating recirculation loop 338 may be made fromhigh-melting-point plastics and/or metal components to resistdeformation and/or promote thermal transfer during the recirculation ofheated fluid to melt and dispense the structural material 326.

The single-use dispensing device 320 may be pre-sterilized and providedwithout any fluid present in the fluid tubing lines. The tubing linesmay then be filled with thermally-regulated fluid prior to use. Thethermally-regulated tubing lines may be filled, properly vented, andrecirculated using an external pump (not shown) and/or pneumaticpressurized source (not shown). The temperature regulation device may bean external device that heats and/or cools the thermally-regulated fluidwithin a container and recirculates the fluid through thethermally-regulated tubing lines within the dispensing device 320.

In some instances the structural material 326 may be partially cooledafter it passes through the heating element 336 to achieve the desireddispensing rate and consistency of the structural material 326 prior toit being dispensed from the structural material dispensing tip 348,particularly if the temperature regulation of the heating element usinga heated fluid is not tightly controlled. The cooling element 342 maycomprise an assembly that circulates a cooling fluid, such as cooled,cold, or chilled sterile filtered water, brine, glycol, and/or air, froman external temperature-regulated fluid source through the cooled tubingline 346. The cooled tubing line 346 may be connected to an internalcooling recirculation loop 344 that may be insulated to protect theinternal components and other temperature controlled fluid lines frombeing negatively affected by the cool temperatures. The cooling element342, the cooled tubing line 346, and the internal cooling recirculationloop 344 may be made from plastics and/or metal components.

Temperature sensors may be embedded into the heating and coolingelements 336, 342 to provide feedback to a computer control board and/oran operator on the dispensing conditions within the dispensing device320. A biological material dispensing element 356 may contain a thermalbarrier to prevent the heating element 336 and/or the cooling element342 and internal thermal fluid control lines 338, 344 from negativelyimpact the dispensing of the biological materials.

The biological material dispensing element 356 may contain a warmingelement 364 to maintain the cells and/or biological material at aconsistent temperature, such as incubation temperature 37′C, duringdispensing. The warming element 364 may comprise an assembly thatcirculates a warm fluid, such as warm sterile filtered water, brine,glycol, and/or air, from an external temperature-regulated fluid sourcethrough the warm tubing line 366. The warming tubing line 366 may beconnected to an internal warming recirculation loop 365 that may beinsulated to protect the internal components and other temperaturecontrolled fluid lines from being negatively affected by the warmtemperatures. The internal warming recirculation loop 365 may bepositioned so that the tubing lines pass through the thermal barrier354. The warming element 364 may be positioned so that it providesconsistent warming and thermal transfer over the entire length of aninternal tubing 362 prior to dispensing out of a biological materialdispensing tip 368.

The biological material dispensing element 356 may be connected to abiological feed source via at least one aseptic connection 358. A ventfilter 360, such as an integrity-testable, sterilizing-grade ventfilter, may be utilized to properly vent the internal tubing 362 as itfills with material. The aseptic connection 358 may be made using anaseptic connector such as an OPTA® connector and/or other physicalaseptic connector, and/or may be achieved by thermowelding a tubinglength of thermoweldable tubing to the feed assembly for a sterileconnection with the biological feed source.

An operator may control the dispensing of the structural and biologicalmaterials through dispense and retraction buttons 350 and a materialselector switch 352 that may switch from dispensing the structuralmaterial 326 to dispensing the biological material. In this example thecontrols may regulate the direction and force of the pneumatic pressurethat controls the flow of materials within the single-use device 320.

FIG. 4C is a side view of a single-use dispensing device 370 comprisinga plurality of single-use chemical heating elements. The single-usedispensing device 370 may be a possible embodiment of the single-usedispensing device 300.

The single-use dispensing device 370 may comprise a printer body 372that encloses a plurality of components manufactured from sterilizablematerials and may be sterilized utilizing a validated method.

The device 370 may comprise a single-use chemical heating element, whichis based on mechanisms such as the exothermic oxidation of iron whenexposed to air, or a re-usable chemical heater element, which is basedon mechanisms such as the exothermic crystallization of supersaturatedsolutions.

A structural material dispensing element 374 of the device 370 maycomprise a structural material 376, a feed motor 378, a venting filter380, internal tubing, a two-chamber chemical heating element 382, 384,and a dispensing tip. The feed motor 378 may be driven by an externalpneumatic pressure source attached to a fluid line 383. The waste air,after running through the feed motor 378, may simply bleed the pressureoff through the venting filter 380 in a direction away from the printedobject or the venting filter 380 may be connected to a length of tubingthat removes the excess compressed air away from the work area.

The feed motor 378 may turn a plurality of gears that feed thestructural material 376 into the internal tubing to the two-chamberheating element 382, 384. The two-chamber heating element 382, 384 maycomprise two different chemical materials, such as calcium oxide andwater, which are kept separated by a seal 388. Exemplarily, when theseal 388 is removed by an operator by pulling on the tape containing theseal 388 from a slot 386 in direction 390, the two separated chemicalsin the two-chamber heating element 382, 384 mix. The mixing causes anexothermic reaction resulting in the heating element temperature toincrease. In other examples, a single-stage chemical heating element maybe utilized (not shown), where the seal 388 may be removed to expose thesingle-stage element to air for the exothermic oxidation of iron as theheating source. The chemical heating method utilized must havesufficient heat and time duration to provide consistent thermal energyfor melting the structural material 376 planned for use.

A biological material dispensing element 392 of the device 370 maycontain a thermal barrier 404 to prevent the chemical heating element382, 384 from negatively impacting the dispensing of the biologicalmaterials. The biological material dispensing element 392 may includetwo-chamber warming element 396, 394 that may comprise two differentchemical materials kept separated by a seal 400. Exemplarily, when theseal 400 is removed by an operator by pulling on the tape containing theseal 400 from a slot 398 in direction 402, the two separated chemicalsin the two-chamber warming element 396, 394 mix, causing an exothermicreaction resulting in the warming element temperature to increase. Thewarming element chamber 394 may use a matrix and/or gel to moderate andcontrol the exothermic reaction to prevent excessive heat from damagingthe biological materials as they move through the internal tubing.Alternatively a supersaturated solution of sodium acetate may utilizeexposure to a metal disc, by removing seal 400, which helps initiatenucleation and the crystallization generating heat from the exothermicreaction.

Temperature sensors may be embedded into the heating and warmingelements to provide feedback to a computer control board and/or anoperator on the dispensing conditions within the dispensing device 370.A plurality of heating and warming element chambers may be utilized tokeep the heating over a defined duration depending on the length of timeit takes to complete the processing of the material into a printedobject. Alternatively the operator may remove the used chemical heatingelements and replace them with new elements such as in a replaceablecartridge configuration. Alternatively the operator may dispose of theprevious dispensing device 370 and connect it to a new dispensing deviceif the chemical heating elements did not last for the duration requiredto complete the printing of the object.

Both the structural material and the biological material dispensed bythe dispensing devices described above may require some kind ofpre-processing prior to being dispensed. Examples of pre-processing mayinclude but are not limited to mixing, selecting, changing theconcentration of the material. FIGS. 5A-5D show examples of dispensingelements comprising different mechanisms for pre-processing of thematerial to be dispensed. Any of the dispensing devices described withreference to the previous figures may comprise one or more of thesedispensing elements.

FIG. 5A is a side view of an single-use biological material dispensingelement 500 containing a mixing device that comprises a shaft with oneor a plurality of impellers 502, which may connect to an externalmagnetic or geared shaft connection 504 containing a seal. The shaftconnection may be the mechanism to drive the rotation of the shaft andthe impellers 502.

An internal tubing for the biological material may contain a pluralityof baffles 506 to aid in the mixing and dispensing of biologicalmaterials aseptically received from a feed source 508. The mixing deviceand baffles 506 may allow for proper mixing and uniformity of aheterogeneous material, for easier dispensing of a viscous material,and/or for promoting the dispensing of either denser or less densematerials such as dispensing cells while removing cell debris.Additionally or alternatively a sparger (not shown) and/or amicrosparger (not shown) with a sterile air connection (not shown) maybe utilized for proper aeration and mixing of the biological materialprior to dispensing. Additionally, the biological materials may becombined with gels, hydrogels, alginates, or thickening agents to aid inthe precision dispensing and attachment to structures and/orscaffolding. The thickening agents may be added through an upstreamconnection of the feed material or within the internal tubing of thesingle-use biological dispensing element 500 via an alternate asepticconnector (not shown). The thickening agents may require vigorous mixingutilizing the mixing device to achieve the proper consistency fordispensing.

FIG. 5B is a side view of a single-use biological dispensing element 510containing one or a plurality of internal roller elements 512 that maybe connected to an external drive mechanism and/or a drive mechanism ofa connected multi-use structural material dispensing element throughmagnetic or geared connections 516. The drive mechanism (not shown) mayutilize a geared shaft, a magnetic assembly, a pneumatic air source,and/or a hydraulic fluid source to drive the internal roller elements512. The internal roller elements 512 may promote mixing and dispensingof biological materials aseptically received from the feed source 514.The roller elements may contain a plurality of protrusions or impellerblades at different pitches and lengths to assist with moving orselecting material with defined sizes prior to dispensing. The internalroller elements 512 may additionally promote the dispensing of specificmaterials such as cells that have a certain defined shape and density.

FIG. 5C is a side view of a single-use biological dispensing element 520containing a one or a plurality of internal membrane elements 522 forthe concentration or dilution of the biological material prior todispensing. The internal membrane elements 522 may be incorporated intoan internal tubing and be utilized to remove fluid material from thebiological material prior to dispensing using an external connection 524and a vacuum force 526. The internal membrane elements 522 may be sizedto properly remove selected materials and to have the force of theremoval controlled to protect the remaining biological materials, suchas for the removal of fluids for concentration while maintaining healthycells for dispensing. In other examples, sterile fluid may be injectedinto the dispensing element 520 through a sterilizing grade membraneelement to dilute the biological materials aseptically received from afeed source 528 to a certain concentration. Examples are for reducingthe trypsin concentration used to remove cells from adherent plates, orto provide a chemical aid for promoting the adhering of the cells orbiological material to the printed structural material. A biologicalmaterial dispensing tip 530 may contain a Luer Lock connection or otherattachment mechanism to attach to a sized filter device 532 that may beplaced just prior to dispensing the biological material. The sizedfilter device 532 may provide sterilizing-grade fluid, thereby removingany potential bacterial contamination while allowing small biologicalmaterials such as proteins to pass through. The plurality of internalmembrane elements 522 and the sized filter device 532 may be integritytestable.

FIG. 5D is a side view of a single-use biological dispensing element 540containing a single-use cell concentrator device 542, such as aHydrocyclone® for the controlled cell retention and concentration priorto dispensing. The single-use cell concentrator device 542 may beaseptically connected to a biological material feed source 544 with twotubing lines. The material is concentrated by means of the flow ofmaterial within the interior of the cell concentrator device 542 throughsedimentation in a centrifugal field, which is similar to centrifugationbut with no movable parts. The cells are concentrated and enter theinternal tubing via a concentrated underfiow connection 546 of thesingle-use biological material dispenser 540, while the cell debris andused media are removed via an overflow connection 548. In otherexamples, the opposite products may be desired and the single-useconcentrator device 542 connections may be changed to supply thesingle-use biological material dispenser 540 with tubing line 548 whilediscarding the material from the concentrated underflow connection 546.In other words, the cells may be discarded and the used media containingthe desired biological material may be harvested and used within theinternal tubing of the single-use biological material dispenser 540.

A handheld dispensing device may be connected to a biological materialsource via an aseptic connection assembly, which may comprise, but isnot limited to, any combination of a filtration train, a surge vesselcontainer, tubes, a fermenter, a cross flow assembly, a membraneadsorber, a centrifugation apparatus, and an incubator.

FIG. 6 shows an example of a system 600 comprising a handheld dispensingdevice 624 and a biological material source 602. The handheld dispensingdevice 624 may comprise the features of one or more of the dispensingdevices described with reference to FIGS. 1A to 5D.

The sterilized printing and feed material system 600 may comprise thebiological material source 602 such as a single-use bioreactor, whichmay be e.g. a microbioreactor, which is connected to a filtration trainvia an aseptic connector 604. The filtration train (or filter train) maycomprise one or a plurality of filters including but not limited to adepth filter 606, a pre-filter 608, and a sterilizing-grade filter 610.The filter train is optional if cells and/or cell products that would becaptured within the filters are the desired biological feed material forprinting.

The filter train may be connected to a surge vessel container 614exemplarily with an additional aseptic connector (not shown) or may besterilized as a complete assembly. The surge vessel container 614 mayfill with the material filtered from the bioreactor 602, which can bedriven by a constant pressure or a constant flow source. A sterilizinggrade vent filter 616 may allow the surge vessel container 614 to ventduring filling. After the filtration process is complete or the surgevessel container 614 is full, a valve 612 to the filter train may beclosed and a regulated compressed air line 618 may be attached to thesterilizing grade air filter 616.

The pressure drives the liquid up a dip tube 620 and into a tubing piecewhich is connected via an aseptic connector 622 to the dispensing device624. The material from the surge vessel container 614 can beconcentrated utilizing a cell retention and concentration device, suchas a gamma irradiatable Hydrocyclone (not shown). Additionally, thebiological material may be connected to a sensor device and/or have anintegrated sensor device for cell counting and cell viability. Thesensor device for cell counting and cell viability may include but isnot limited to a nucleocounter, a flow cytometer, and/or a radiofrequency impedance device, preferably a BioPat® ViaMass Sensor unit.

An operator may utilize the dispensing device 624 within a cleanenvironment with sufficient airflow to prevent contamination, such as alaminar flow hood or a biological safety cabinet, or within a sterilizedenvironment such as an isolator, glovebox, or sterile chamber. Theoperator may precisely control the dispensing of the structural materialonto a printing tray or container such as a multi-well plate 626 usingthe structural material dispensing element of the dispensing device 624to form a scaffolding or structure to support the biological materials.The processed biological material may be precisely deposited using thebiological material dispensing element of the dispensing device 624 ontothe structural material within the multi-well plate 626. The 3D printedobject within the multi-well plate 626 is formed by layer-by-layeradditive printing of structural material from the dispensing device 624that is manually operated. After the printing of the structuralmaterial, the biological material may be added and the printed objectwithin the multi-well plate 626 may be filled with a nutrient fluidmedia. The multi-well plate 626 may be covered, removed from theprinting environment, and incubated within an incubated container forfurther study, sampling, and/or evaluation.

FIG. 7 shows an example of a system 630 comprising a handheld dispensingdevice 680 and a plurality of biological material sources, such as twosingle-use bioreactors 632, 654. The handheld dispensing device 680 maycomprise the features of one or more of the dispensing devices describedwith reference to FIGS. 1 to 5.

The sterilized printing and feed material system 630 may comprise atleast two single-use bioreactors 632, 654 that are connected to theirrespective filtration assemblies via aseptic connectors 634, 656. Thefiltration train for the first single-use bioreactor 632 may compriseone or a plurality of filters including but not limited to a depthfilter 636, a pre-filter 638, and a sterilizing-grade filter 640. Thefiltration train for the second single-use bioreactor 654 may compriseone or a plurality of filters including but not limited to a pre-filter658, a mycoplasma retentive filter 660, and a virus retentive filter662. The filter train assemblies are optional if cells and/or cellproducts that would be captured within the filters are the desiredbiological feed material for printing.

The filter train assemblies may be connected to surge vessel containers644, 666 exemplarily with the use of additional aseptic connectors (notshown) or may be sterilized as complete assemblies. The surge vesselcontainers 644, 666 may fill with the material processed from thesingle-use bioreactors 632, 654, which can be driven by a constantpressure or a constant flow source. Sterilizing grade vent filters 646,668 may allow the surge vessel containers 644, 666 to vent duringfilling. After the filtration processes have completed or the surgevessel containers 644, 666 are full, valves 642, 664 to the filter trainassemblies may be closed and regulated compressed air lines 648, 670 maybe attached to sterilizing-grade air filters 646 and 668 respectively.

The pressure drives the liquid up the dip tubes 650, 672 and into tubingpieces that are connected via aseptic connectors 652, 674 to thedispensing device 680. A plurality of feed sources of biologicalmaterials may be aseptically connected to a manifold 676 on thedispensing device 680. A selection valve 678 located on the manifold 676may be utilized to select the feed source to be connected to theinterior tubing of the single-use biological material dispensing elementduring dispensing.

An operator may precisely control the dispensing of the structuralmaterial onto a printing tray or container such as a multi-well plate682 using the structural material dispensing element of the dispensingdevice 680 to form a scaffolding or structure to support the biologicalmaterials. The processed biological material may be precisely depositedusing the biological material dispensing element of the dispensingdevice 680 onto the structural material within the multi-well plate 682.The three dimensional printed object within the multi-well plate 682 isformed by layer-by-layer additive printing of structural material fromthe dispensing device 680 manually operated. After the printing of thestructural material, the biological material may be added and theprinted object within the multi-well plate 682 may be filled with anutrient fluid media. The dispensing device 680 may dispense materialinto at least one well within the multi-well plate 682 from a singlebiological feed source such as the material originating from asingle-use bioreactor 632 or the material originating from a pluralityof single-use bioreactors 632, 654 may be dispensed within at least onewell within the multi-well plate 682. The different feed sources may bedispensed by the operator onto different sections of the structuralmaterial or scaffolding to form a complete object. The multi-well plate682 may be covered, removed from the printing environment, and incubatedwithin an incubated container for further study, sampling, and/orevaluation.

FIG. 8 shows an example of a system 700 comprising a handheld dispensingdevice 742, a biological material source such as a single-use bioreactor702, a centrifugation assembly 706 and a crossflow assembly 728. Thehandheld dispensing device 742 may comprise the features of one or moreof the dispensing devices described with reference to FIGS. 1 to 5.

The sterilized printing and feed material system 700 may comprise asingle-use bioreactor 702 connected to the centrifugation assembly 706via an aseptic connector 704. The filtration train assembly may beconnected to the centrifugation assembly 706 via an aseptic connector(not shown) and comprise one or a plurality of filters including but notlimited to a depth filter 708, a pre-filter 710, and a sterilizing-gradefilter 712. The filter train is optional if cells and/or cell productsthat would be captured within the filters are the desired biologicalfeed material for printing.

The filter train assembly may be connected to a surge vessel container716 exemplarily with an additional aseptic connector (not shown) or maybe sterilized as a complete assembly. The surge vessel container 716fills with the material filtered from the bioreactor, which can bedriven by a constant pressure or a constant flow source. A sterilizinggrade vent filter 718 may allow the surge vessel container 716 to ventduring filling. After the filtration process is complete or the surgevessel container 716 is full, a valve 714 to the filter train may beclosed and a regulated compressed air line 720 may be attached to thesterilizing grade air filter 718.

The pressure drives the liquid up a dip tube 722 and into a tubing piecewhich may be connected via an aseptic connector 722 to a pre-sterilizedmembrane adsorber 726. The membrane adsorber 726 may be achromatographic membrane carrying functional groups for the reversiblebinding of biomolecules. The desired molecules can be captured with themembrane adsorber and eluted at a later time or undesirable moleculescan be removed by membrane adsorption before further processing. Themembrane adsorber 726 may be connected to a pre-sterilized cross flowassembly 728. The cross flow assembly 728 may comprise a plurality ofmicrofiltration and/or ultrafiltration cassettes in varying sizes.

The cross flow assembly 728 may be connected to a surge vessel container732 via an aseptic connector (not shown). The surge vessel container 732fills with the material filtered and/or concentrated from the cross flowassembly 728, which may be driven by a constant pressure or a constantflow source. Additionally or alternatively, other common bioprocessprocessing methods of the biological material may occur, such as columnchromatography, high performance liquid chromatography (HPLC), fastprotein liquid chromatography (FPLC), and one of these other processesmay fill the surge vessel container 732. A sterilizing grade vent filter734 may allow the surge vessel container 732 to vent during filling.After the cross flow processing is complete or the surge vesselcontainer 732 is full, a valve 730 to the cross flow assembly may beclosed and a regulated compressed air line 736 may be attached to thesterilizing grade air filter 734.

The pressure drives the liquid up a dip tube 738 and into a tubing piecethat may be connected via an aseptic connector 740 to the dispensingdevice 742. An operator may precisely control the dispensing of thestructural material onto a printing tray or container such as amulti-well plate 744 using the structural material dispensing element ofthe dispensing device 742 to form a scaffolding or structure to supportthe biological materials. The processed biological material may beprecisely deposited using the biological material dispensing element ofthe dispensing device 742 onto the structural material within multi-wellplate 744. The 3D printed object within the multi-well plate 744 isformed by layer-by-layer additive printing of structural material fromthe dispensing device 742 manually operated. After the printing of thestructural material, the biological material may be added and theprinted object within the multi-well plate 744 may be filled with anutrient fluid media. The multi-well plate 744 may be covered, removedfrom the printing environment, and incubated within an incubatedcontainer for further study, sampling, and/or evaluation. Additionallyor alternatively the operator may dispense the biological material ontoan alternate substrate, such as a membrane (not shown) and/or diagnosticstrips (not shown). The dispensing device 742 may spray deposit proteinsand/or other concentrated ultra-filtered materials onto the membranesstrips for use in diagnostic analysis. Additionally other structuralcomponents may be added to the membrane strips by layer-by-layeradditive printing of material from the dispensing device 742.

As described with reference to FIGS. 1 and 2, the dispensing device maycomprise positional sensors. Generally, positional sensors may be formedwithin, connected to, or inserted within the dispensing device fortracking the orientation and movement in a three dimensional space. Thedispensing device may additionally comprise other internal sensors tocollect additional information on the dispensing process, such as volumedispensed or dispensing speed. Alternatively, data on the dispensingprocess of the dispensing device such as position, orientation,movement, materials dispensed, volume dispensed, and conditions may betracked within a workspace using an external monitoring device. The datagenerated from the internal sensors and/or external monitoring devicemay be stored as a file in an internal memory storage device, anexternal memory storage device, a connected memory storage device,and/or a networked memory storage device. The stored data may be edited,e.g. scaled-up, and used to print a plurality of replicas of a 3D objectwithin a large-scale 3D printer, such a single-use biological 3Dprinter.

FIG. 9 shows an example of an external monitoring setup for themonitoring of a handheld dispensing device 1010. The handheld dispensingdevice 1010 may comprise the features of one or more of the dispensingdevices described with reference to the previous figures.

FIG. 9 shows a front view of an external monitoring system setup 1000including an augmented reality system 1006 that comprises a camera andlighting array sensing device 1002 with a wired/wireless connection to adisplay device 1004. The camera and lighting array sensing device 1002may contain one or a plurality of cameras, such as video cameras, depthcameras, infrared cameras, thermal cameras, or Light detection andranging (LIDAR), etc. as well as one or a plurality of adjustable lightsto fully illuminate the workspace for optimal tracking of markers. Thecamera and lighting array sensing device 1002 may monitor and image aworkspace 1008, which may contain one or a plurality of identificationmarkers.

The workspace 1008 may be within a clean environment with sufficientairflow to prevent contamination, such as a laminar flow hood or abiological safety cabinet, or within a sterilized environment such as anisolator, glovebox, or sterile chamber. The workspace 1008 mayadditionally be on a benchtop or desk in a non-sterile or non-cleanenvironment for practicing printing a plurality of three dimensionalobjects following a defined protocol.

The one or more identification markers may identify objects within theworkspace field of view and provide linked information on them, codedprotocol identifiers to load a specific program of instructions from adatabase to be followed for printing a plurality of objects. Theidentification markers may for e.g. be static or variableaugmented-reality markers. An augmented-reality marker may be a physicalor virtual tag that provides unique identification information andpositional information. A variable augmented-reality marker may have atleast two states (e.g. it shows two different images) and thepresentation of one of those states may be triggered by an operator'sinput or a computer product input, e.g. at a programmed interval.Coordinate markers 1022, 1024 may provide an augmented coordinate system1026 where objects between the coordinate markers 1022, 1024 and withinthe field of view of the sensing device 1002 may be tracked based on therelative distance between the coordinate markers 1022, 1024, which maybe viewed as an augmented image on the display device 1004 as describedin U.S. Pat. No. 8,749,396 B2, which is incorporated herein byreference. The display device 1004 may be a monitor, screen, orprojection display, or a wearable display such as a head mounted displaydevice (HMD), an augmented reality display, a virtual reality display,or a mixed reality display device.

An operator 1012 may use the dispensing device 1010 to dispensestructural materials and/or biological materials into a printer tray ordispensing container, which for example may be a multi-well plate 1020.The augmented reality system 1006 can precisely track the movement ofthe dispensing device 1010 within the workspace 1008 in three dimensionsthrough the use of one or a plurality of light emitting diodes 1018,which may exemplarily be infrared light emitting diodes and wherein eachinfrared LED emits a different specific wavelength, and a variablemarker 1016 both located on the dispensing device 1010. The variablemarker 1016 on the dispensing device 1010 can change the presentation tothe augmented reality system 1006 depending on the material selected tobe dispensed, the initiation of dispensing, the rate of dispensing, theconditions of dispensing, and the volume of material dispensed. Theaugmented reality system 1006 may present an augmented display 1028 of aplurality of 3D virtual objects 1032, showing the material types, thematerial shapes, and the material volumes required to be dispensed intoeach well of the multi-well plate 1020 or printing tray in accordancewith a defined protocol.

The augmented reality system 1006 may record the movement of theinfrared LED 1018 as well as the location and presentation of thevariable marker 1016 as a 3D wireframe diagram, which is a virtual linethat tracks the movement of the marker in three-dimensional space overtime, and compares the movements of the marker with a reference diagramwithin a pre-programmed tolerance, as described in U.S. Pat. No.8,982,156 B2, which is incorporated herein by reference. As the variablemarker 1016 changes its presentation to the augmented reality system1006, the changes may be recorded on the wireframe diagram stored by theaugmented reality system. The dispensing device 1010 may comprise one ora plurality of positional sensors 1014 including but not limited tomotion sensors, orientation sensors, gyroscopic sensors, environmentalsensors, cameras, depth sensors, magnetometers, accelerometers,proximity sensors, GPS devices, IMUs and internal or externalpositioning sensors. The information from the one or more orientationand positioning sensors of the dispensing device assembly 1010 may becommunicated to the augmented reality system 1006 via a wired orwireless connection to provide additional real-time positionalinformation for the augmented reality display 1004.

The augmented reality system 1006 may additionally provide asuperimposed video and/or animation 1030 of the correct procedure andproper positioning of the dispensing device 1010 to physically print a3D virtual object 1032. The procedure may be performed on the display1004 so that the operator can follow the superimposed video and/oranimation 1030 to correctly complete the proper procedures and thesequence for the performance of a work task. The superimposed videoand/or animation 1030 may automatically speed up or slow down to mimicthe speed and steps of the operator 1012 based on the infrared LEDmarker tracking, the variable marker tracking, the positional sensortracking, or a combination thereof. Alternatively, the positionaltracking information of the dispensing device 1010, the augmenteddisplay 1028 and the 3D virtual object 1032 to be printed may bedisplayed on a virtual reality display device (not shown) or mixedreality display device (not shown). The operator may be evaluated andgraded by the software that tracks the movements and presentation of thevariable markers within an operator-defined system for performing orfollowing the proper technique for a predetermined task within themargin of error. If the operator meets a certain proficiency level, theaugmented reality system 1006 may qualify the operator as meeting therequirements for performing the work task. This qualification can be theinitial qualification for the operator or one of pre-programmed periodicqualifications, such as part of an annual re-training and evaluation.

The dispensing device shown in FIGS. 1 to 9 is a handheld dispensingdevice. However, such a dispensing device, i.e. with the samecharacteristics, may alternatively be controlled by a robotic controltool. According to one example, the robotic control tool may be manuallycontrolled by an operator via a communication with an external controldevice maneuvered by the operator. According to another example, therobotic control tool may be controlled by an automatically-operatedexternal control device and run a pre-programmed operation for printing.FIGS. 10 and 11 show examples of a dispensing system comprising asterilizable chamber, a dispensing device and an external controldevice.

The dispensing system 800 shown in FIG. 10 may include a sterilizablechamber 802 that comprises at least one robotic arm assembly 804, a port814 to connect the external control device 830 with the robotic armassembly 804, and a transfer hatch 810 for the chamber 802. The chamber802 may be within a clean environment with sufficient airflow to preventcontamination, such as a laminar flow hood or a biological safetycabinet, or within a sterilized environment such as an isolator,glovebox, or sterile chamber. In one example, the chamber 802 may be apre-sterilized chamber containing the robotic arm assembly 804, adispensing device 806 and a plurality of printing trays or containerssuch as multi-well plates 808. The dispensing device 806 may be asingle-use assembly that has been sterilized along with thepre-sterilized chamber 802 or it may be sterilized separately using analternative sterilization method from the sterile chamber andaseptically inserted utilizing an aseptic connection and insertionmethod (not shown).

Exemplarily, the robotic arm assembly 804 may utilize one or a pluralityof actuators to control the positioning and movements of the dispensingdevice. The robotic arm assembly 804, including the plurality ofactuators, may be made out of sterilizable (gamma irradiatable and/orautoclavable) plastic materials and rubberized seals. For example, therobotic arm assembly 804 may be controlled by utilizing an actuator torotate a rotating base. The actuators may be driven by hydraulic,pneumatic, electric, or magnetically controlled methods (not shown). Inone example, the movements of the robotic arm assembly 804 may becontrolled via an external control device, which may be a manualcontroller operated by an operator external to the sterilized chamber802.

The external control device 830 may utilize one or more hydraulic and/orpneumatic tubing lines 818 to move fluid through the port 814 into theactuators located on the robotic arm assembly 804 to control themovements. The tubing lines 818 may be filled with a sterile fluidthrough a filling assembly 820, wherein the fluid may apply hydraulicand/or pneumatic pressure. For example, the tubing lines may be filledwith a sterile hydraulic fluid such as sterile filtered water. Thesterile filtered water may enter into the robotic arm assembly 804 andthe external control device 830 after sterilization and setup for use byan operator. The tubing lines to the internal robotic arm assembly 804may be connected to the external filling assembly 820 and/or theexternal control device 830 utilizing an aseptic connector (not shown).Purified water may enter through a tubing line 824, pass through asterilizing grade filter 826 and then enter into the filling assembly820, which may serve as a manifold to completely fill each of theindividual tubing lines 818. The interior of the tubing lines 818 may becleared of air through a sterilizing grade vent filter 828 that allowsthe displaced air to vent to the atmosphere as it is displaced bysterile filtered water entering into the assembly. A valve 822 on thefilling assembly 820 may be closed when the charging of the fluid lineshas been completed. Additional sterile filtered water, other fluids, orin other examples air or gas may be added to the tubing lines 818 viathe filling assembly 820 in case of leakage or loss of pressure.

The external control device 830 may be utilized to control the movementsof the robotic arm assembly 804. The external control device 830 may bein the shape of the dispensing device 806 to simulate the movement andcontrol of the device by the operator's hand 834. Alternatively theexternal control device 830 may be in the shape of a stylus or otherminimalized simulant of the actual shape or design of the dispensingdevice 806, which provides movement and external control by the operatorof the robotic arm assembly 804 internal to the sterile chamber 802.

A plurality of hydraulic and/or pneumatic pistons may be arranged as apiston assembly 836 and filled with fluid, wherein the pistons may bepushed and/or pulled by the operator. This movement of one of thepistons 836 may alter the movement of a seal internal to the piston andalter the displacement of the internal fluid, resulting in the movementof the corresponding pistons 812 on the robotic arm assembly 804. Themovement is transmitted by the pressure of the fluid that moves throughthe tubing lines 818 into the sterile chamber 802.

A body 832 of the external control device 830 may contain all of thesame buttons, switches, and displays as the dispensing device 806. Adata cable 838 may transmit the data resulting from e.g. the pushing ofa button or other inputs from the body 832 to the internal dispensingdevice 806 through the corresponding internal data cable 816. The datacables 816, 838 may provide data, power, and/or fluid communication tothe internal dispensing device 806. Exemplarily, commands from theexternal control device may include but are not limited to thedispensing of material using the dispense and retraction buttons and theselection of which material (such as the structural material or thebiological material) to dispense using the material selector switch.Conversely the internal data cable 816 may transmit sensor data andrelated information, such as position and orientation, from the internaldispensing device 806 to the corresponding data cable 838 on theexternal control device body 832.

The internal dispensing device 806 and/or the external control device830 may contain a plurality of positional sensors including but notlimited to motion sensors, orientation sensors, gyroscopic sensors,environmental sensors, cameras, depth sensors, magnetometers,accelerometers, proximity sensors, GPS devices, IMUs and internal orexternal positioning sensors. The information from the orientation andpositioning of the biological three dimensional material dispensingdevice assembly 806 used by an operator can be saved into a file whichmay be utilized to print out a replica of the structural materials andbiological materials in the exact same way as was manually printedutilizing a large-scale single-use biological 3D printer (not shown).

The purpose of the robotic arm assembly 804 is to move, manipulate, anddispense materials from the dispensing device 806 internal to thesterilized chamber 802 while the operator is operating comfortablyoutside of the sterilized chamber 802. Indeed, operating within anisolator or glovebox typically means dealing with bulky gloves, so thatthe fine controls of dispensing materials into a small printing tray ormulti-well plate 808 may not be adequately maneuvered.

The transfer hatch 810 may be utilized to remove objects asepticallyfrom the sterilized chamber 802 and/or to connect to additionalsterilized chambers to form a more complex assembly. The robotic armassembly 804 may additionally be utilized to move materials from onesterilized chamber to another through the connected transfer hatches810.

FIG. 11 shows a dispensing system 850 comprising a chamber 852 that mayinclude at least one robotic arm assembly 854, a port 865 to connect toan automated control assembly (or automatically-operated externalcontrol device) 880, and a transfer hatch 860 for the chamber 852. Thechamber 852 may be within a clean environment with sufficient airflow toprevent contamination, such as a laminar flow hood or a biologicalsafety cabinet, or within a sterilized environment such as an isolator,glovebox, or sterile chamber. Exemplarily, the chamber 852 may be apre-sterilized chamber containing the robotic arm assembly 854, adispensing device 856 and a plurality of printing trays or containerssuch as multi-well plates 858. The dispensing device 856 may be asingle-use assembly that has been sterilized along with thepre-sterilized chamber 852 or it may be sterilized separately using analternative sterilization method from the sterile chamber andaseptically inserted utilizing an aseptic connection and insertionmethod (not shown).

In one example the robotic arm assembly 854 may utilize a plurality ofactuators to control the positioning and movements of the dispensingdevice 856. The robotic arm assembly 854, including the plurality ofactuators, may be made out of sterilizable (gamma irradiatable and/orautoclavable) plastic materials and rubberized seals. Exemplarily, therobotic arm assembly 854 may be controlled by utilizing an actuator torotate a rotating base. The actuators may be driven by hydraulic,pneumatic, electric, or magnetically controlled methods (not shown).

In one example the robotic arm assembly 854 may be controlled via theexternal automated control assembly 880. The external automated controlassembly 880 may utilize a plurality of hydraulic and/or pneumatictubing lines 866 to move pressurized fluid into an actuator assembly 862located on the robotic arm assembly 854 to control the movements. Thetubing lines 866 may be filled with a sterile fluid through a fillingassembly 868, wherein the fluid may apply hydraulic and/or pneumaticpressure. For example, the tubing lines may be filled with a sterilehydraulic fluid such as sterile filtered water. The sterile filteredwater may enter into the robotic arm assembly 854 and the externalcontrol device 880 after sterilization and setup for use by an operator.The tubing lines to the internal robotic arm assembly 854 may beconnected to the external filling assembly 868 and/or the externalcontrol device 880 utilizing an aseptic connector (not shown). Purifiedwater may enter through a tubing line 870, pass through a sterilizinggrade filter 872 and then enter into the filling assembly 868, which mayserve as a manifold to completely fill each of the individual tubinglines 866. The interior of the tubing lines 866 may be cleared of airthrough a sterilizing grade vent filter 874 that allows the displacedair to vent to the atmosphere as it is displaced by sterile filteredwater entering into the assembly. A valve 876 on the filling assembly868 may be closed when the charging of the fluid lines has beencompleted.

The automated control assembly 880 may be utilized to control themovements of the robotic arm assembly 854. Pluralities of hydraulicand/or pneumatic pistons arranged in a piston assembly 882 may be filledwith fluid and the movements of the piston heads (not shown) may beautomatically controlled with a processing device 886. A memory storagedevice 888 may store programs local to the automated control assembly880 to control the movements of the robotic arm assembly 854 within aprogram for executing a printing of an object within the sterile chamber852 using the robotic arm assembly 854 and the internal dispensingdevice 856. The processing device 886 may also process sensor data andalter the movements of the robotic arm assembly 854 based on theprograms stored within the memory storage device 888.

A data cable 864 from the dispensing device 856 may connect to theautomated control assembly 880 through data connection 884 tocommunicate data from the pre-programmed button pushes and settings atthe automated control assembly 880 and send the data to the internaldispensing device 856 through the data cable 864. The data cable 864 mayprovide data, power, and/or fluid communication to the internaldispensing device 856 from the data connection 884 on the automatedcontrol assembly 880. Examples of commands from the automated controlassembly 880 include but are not limited to the dispensing of materialand the selection of which material to dispense. Conversely, theinternal data cable 864 may transmit sensor data and relatedinformation, such as position and orientation, from the internaldispensing device 856 to the corresponding data connection 884 on theautomated control assembly 880.

The automated control assembly 880 may additionally contain a wirelesscommunication device 890 to import robotic arm assembly controlprotocols, wireless sensor data, through communication with an externalnetwork and/or a mobile device. The automated control assembly 880 mayutilize the wireless communication device 890 to receive wireless data902 from a wireless external input controller 894, which is a manualcontroller operated by an operator 895 external to the sterilizedchamber 852. The wireless external input controller 894 may utilizepositional information to send instructions to the automated controlassembly 880 to control the movements of the robotic arm assembly 854internal to the sterilized chamber 852. The wireless external controldevice 894 may be in the shape of the dispensing device 856 to simulatethe movement and control of the device by the operator's hand 895.Alternatively the wireless external control device 894 may be in theshape of a stylus or other minimalized simulant of the actual shape ordesign of the dispensing device 856.

The wireless external control device 894 may contain a processing device896, a memory storage device 898, a power device (not shown), such as anelectrically-wired source or battery-powered source, and a communicationdevice 900 to wireless communicate 902 the data from the movement of thewireless external control device 894 to the automated control assembly880 or to another computer device. The wireless external control device894 may contain a robotic arm assembly stand 892 that mimics the roboticarm assembly in the sterile chamber to provide the operator with thefeeling of controlling the actual device.

The wireless external control device 894 may contain all of the samebuttons, switches, and displays as the dispensing device 856. Thecommands from the wireless external control device 894, including thedispensing of material using the dispense and retraction buttons and theselection of which material to dispense using the material selectorswitch, may be wirelessly communicated 902 to the automated controlassembly 880 or another computer device. Data from sensors in theactuators on the robotic arm assembly stand 892 may be communicated tothe wireless external control device 894 for processing, memory storage,and external communication. The wireless data 902 from the wirelessexternal control device 894 may be utilized for real-time printingwithin the sterile chamber 852 by the movements of the robotic armassembly 854 and dispensing with the internal dispensing device 856.Alternatively the wireless data 902 from the wireless external controldevice 894 may be stored in a memory device 888, 898 as an object printfile for execution at a later time.

The internal dispensing device 856 and/or the wireless external controldevice 894 may contain a plurality of positional sensors including butnot limited to motion sensors, orientation sensors, gyroscopic sensors,environmental sensors, cameras, depth sensors, magnetometers,accelerometers, proximity sensors, GPS devices, IMUs and internal orexternal positioning sensors. The information from the orientation andpositioning of the dispensing device 856 or the wireless externalcontrol device 894 used by an operator can be saved into a file whichmay be utilized to print out a replica of the structural materials andbiological materials in the exact same way as was manually printedutilizing a large-scale single-use biological 3D printer 910.Additionally or alternatively the data communicated 902 from thepositional sensors on the wireless external control device 894 may beutilized in real-time for practice printing virtual objects utilizing anaugmented reality display 904, a virtual reality display 906, and/or amixed reality display (not shown). The operator may move the wirelessexternal control device 894 and the comparable movements and dispensingactions may be enacted virtually through a computer program usingaugmented reality, virtual reality, and/or mixed reality. This willallow the operator to practice multiple printing of virtual objectsprior to printing an actual object without wasting valuable structuralmaterial and/or valuable biological materials. The files for printing inan augmented reality, virtual reality, and/or mixed reality environmentmay be saved in a memory storage device and executed using the automatedcontrol assembly 880 and/or the large-scale single-use biological 3Dprinter 910.

The purpose of the robotic arm assembly 854 is to move, manipulate, anddispense materials from the dispensing device 856 internal to thesterilized chamber 852 while the operator is operating comfortablyoutside of the sterilized chamber 852. Indeed, operating within anisolator or glovebox typically means dealing with bulky gloves, so thatthe fine controls of dispensing materials into a small printing tray ormulti-well plate 858 may not be adequately maneuvered.

The transfer hatch 860 may be utilized to remove objects asepticallyfrom the sterilized chamber 852 and/or to connect to additionalsterilized chambers to form a more complex assembly. The robotic armassemblies may additionally be utilized to move materials from onesterilized chamber to another through the connected transfer hatches860.

The large-scale single-use biological 3D printer 910 may be operatedwithin a sterile chamber 912 with a printing platform 918 driven by ahydraulically and/or pneumatically linear actuator and a printer head926 featuring a plurality of articulating axis joints 922 and 924 asdescribed in U.S. patent application Ser. No. 14/927,848, which isincorporated herein by reference. The at least two articulating axisjoints 922 and 924 on the printer head 926 allow for the positioning ofa printer head dispenser 928 at a plurality of angles in relation to theprinting tray 920. The hydraulically- and/or pneumatically-driven linearactuators 914 and 916 are extended at different heights resulting in theprinting platform 918 and printing tray 920 to be positioned at an anglein relation to the printer head dispenser 928. The multi-axispositioning of the printing tray 920 at a plurality of angles, as wellas the multi-axis positioning of the printer head dispenser 928, allowsfor additional degrees of freedom and increased flexibility for printingwith a printer setup over a standard 3-axis coordinate printing system.These additional degrees of freedom mimic the movements an operatorwould make during the printing of an object with the dispensing device856 device and/or the wireless external control device 894.

What is claimed is:
 1. A dispensing system comprising: a sterilizablechamber; at least one robotic arm assembly and a dispensing devicewithin the sterilizable chamber, wherein the at least one roboticassembly is configured to move the dispensing device comprising a firstdispensing element and a second dispensing element, the first dispensingelement being connectable to the second dispensing element; and anexternal control device connected to the at least one robotic armassembly and configured to control the at least one robotic armassembly; wherein the first dispensing element is configured to dispensea structural material and the second dispensing element is configured todispense a biological material.
 2. The dispensing system of claim 1,further comprising at least one tubing line filled with a sterile fluid,the at least one tubing line configured to control the movements of theat least one robotic arm assembly by means of the pressure exercised bythe sterile fluid.
 3. The dispensing system of claim 1, wherein theexternal control device is manually operable.
 4. The dispensing systemof claim 1, wherein the external control device is an automated controlassembly comprising a wireless communication device and the dispensingsystem further comprises a wireless external input controller incommunication with the wireless communication device.
 5. The dispensingsystem of claim 4, further comprising a computing device and wherein thewireless external input controller further comprises positional sensorsand is configured to communicate data from the positional sensors to thecomputing device, wherein the computing device is configured tovirtually replicate movements of the wireless external input controller.